Construction of Isaria fumosorosea blastospore-transforming system by agrobacterium-mediated transformation with benomyl-resistance gene

The green fluorescent protein (gfp) and benomyl resistance gene,the former biomarker for the study of host-pathogen interactions,has been expressed in a variety of prokaryotic and eukaryotic organisms. here we report the expression of gfp in the entomopathognic fungus isaria fumosorosea through agrobacterium-mediated transformation (atmt) using the pk-ben-gfp vector that confers resistance to benomyl. all transformants expressed gfp whose signal was readily detected by fluorescence microscopy. furthermore,fluorescent hyphae and conidia were easily distinguished on insect host,plutella xylostella,and blastospores were also detected in the hemolymph of the diamondback moth larvae. our results show that gfp-tagged strains of i. fumosorosea can be used to study the developmental fate of the fungus within its insect hosts and for analysis of the expression of tagged genes. at the same time,genetic transformation of isaria with benomyl resistant gene indicates that these fungi can be used in combination with some fungicides to protect crop from insects as well as plant disease. copyright 2015 zoological society of pakistan.